Department of Medicine II, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Methods Mol Biol. 2024;2837:219-226. doi: 10.1007/978-1-0716-4027-2_19.
HBV-specific CD8+ T cells are only present at the low frequency during chronic infection. Thus, they are often undetectable by conventional ex vivo staining methods using peptide-loaded HLA class I tetramers. Detection sensitivity can be increased by magnetic bead-based enrichment strategies following staining with peptide-loaded HLA class I tetramers. Additionally, some downstream applications like e.g., single cell RNA sequencing of virus-specific CD8+ T cells may also require a pre-enrichment step to increase the frequency of the cells of interest. For this, peptide-loaded HLA class I tetramers-associated magnetic bead-based enrichment is also a suitable method.
在慢性感染期间，HBV 特异性 CD8+ T 细胞的频率很低。因此，它们通常无法通过使用负载肽的 HLA Ⅰ类四聚体的传统体外染色方法检测到。通过在用负载肽的 HLA Ⅰ类四聚体染色后进行基于磁珠的富集策略，可以提高检测灵敏度。此外，一些下游应用，例如病毒特异性 CD8+ T 细胞的单细胞 RNA 测序，也可能需要预富集步骤来增加感兴趣细胞的频率。为此，负载肽的 HLA Ⅰ类四聚体相关的基于磁珠的富集也是一种合适的方法。
